This research proposal will characterize dominant ovarian antigens recognized by serum antibodies from patients with premature ovarian failure (POF) with the goals of better understanding the pathophysiology of POF and identifying biomarkers for diagnosis of autoimmune ovarian disease. Sera from patients with POF will be screened for antiovarian antibodies and then employed on 2-D gel immunoblots to identify immunoreactive proteins present in human and mouse ovarian and egg extracts. Immunoreactive ovarian and egg proteins will be microsequenced to identify candidate genes encoding ovarian and egg antigens. Peptide microsequences obtained by mass spectrometry will be compared to gene and protein databases to identify autoantigens. Full length cDNAs corresponding to ovarian/egg autoantigens will be cloned and recombinant antigens expressed in bacterial and yeast expression systems. The incidence of antibodies to one or more defined recombinant egg target antigens will then be determined in patients with POF and antiovarian antibodies. The studies are anticipated to lead to a deeper understanding of ovarian autoimmunity in humans and to improved methods of detecting antiovarian antibodies including the possibility of detecting early stage disease. Availability of better diagnostics for ovarian disease may also help in proper clinical follow up and selection of patients who may benefit from immunotherapy or assisted reproductive technologies (in vitro fertilization/embryo transfer [IVF/ET]) to recover their fertility. [unreadable] [unreadable] [unreadable]